Structure-Activity Relationships of Benzamides and Isoindolines Designed as SARS-CoV Protease Inhibitors Effective against SARS-CoV-2

Inhibition of coronavirus (CoV)-encoded papain-like cysteine proteases (PL^pro) represents an attractive strategy to treat infections by these important human pathogens. Herein we report on structure-activity relationships (SAR) of the noncovalent active-site directed inhibitor (R)-5-amino-2-methyl-N-(1-(naphthalen-1-yl)ethyl) benzamide ( 2 b ), which is known to bind into the S3 and S4 pockets of the SARS-CoV PL^pro. Moreover, we report the discovery of isoindolines as a new class of potent PL^pro inhibitors. The studies also provide a deeper understanding of the binding modes of this inhibitor class. Importantly, the inhibitors were also confirmed to inhibit SARS-CoV-2 replication in cell culture suggesting that, due to the high structural similarities of the target proteases, inhibitors identified against SARS-CoV PL^pro are valuable starting points for the development of new pan-coronaviral inhibitors.     

A new method for the evaluation of thermal conductivity and thermal diffusivity from transient hot strip measurements

The standard straight-line fit to data of a transient hot strip (THS) experiment to determine the thermal conductivity and thermal diffusivitya suffers from two major drawbacks: First, due to the statistical nature of the estimation procedure, there is no relation between the uncertainty of the measured value on one hand and the transport properties obtained on the other. Second, in order to account for he heat capacity of the strip and outer boundary conditions, two intervals of the plot must he rejected before analyzing it. So far, these intervals are selected arbitrarily. We now treat the THS working equation as a function of the four parameters concerned. a.U ₀ (initial voltage), andt ₀ (time delay). Chi-square fittings. following the Levenberg-Marquardt algorithm. are performed separately for several overlapping time intervals of the entire plot to find and a with minimal standard deviation. In the course of subsequent iterations an individual weighting factor is applied to each point to account for systematic errors. This procedure yields the "best" values of anda along with their individual errors. comprising the systematic and the statistical errors. Experimental results on Pyrex glass 7740 were taken to verify the new procedure.Paper presented at the Twelfth Symposium on Thermophysical Properties, June 19–24, 1994, Boulder. Colorado, U.S.A.     

Characterization of a High-Temperature Thermal Conductivity Reference Material

The development of thermal conductivity reference materials for high-temperature insulation consists of three stages: provisional assessment of candidate reference materials, detailed assessment of candidate reference material(s) and corresponding inter-laboratory comparisons. This paper describes the detailed characterization of a candidate high-temperature thermal conductivity reference material, a high-density calcium silicate (HDCaSi-N). The selection criteria, assessments of uniformity and stability, the thermal expansion behavior and their effects on the thermal conductivity reference specimens are presented in the paper. The uniformity assessments include the thermal expansion variation in orthogonal orientations and different locations and from different boards, as well as thermal conductivity variation within the batch of the specimens. The dimensional stability assessment in terms of thermal expansion and the short-term stability in terms of thermal conductivity are also presented.     

Design Guideline for New Generation of High-Temperature Guarded Hot Plate

This paper complements the existing measurement standards and literature for high-temperature guarded hot plates (HTGHPs) by addressing specific issues relating to thermal conductivity measurement of technical insulation at high temperatures. The examples given are focused on the designs of HTGHPs for measuring thin thermal insulation. The sensitivity studies have been carried out on major influencing factors that affect the thermal conductivity measurements using HTGHPs, e.g., the uncertainty of temperature measurements, plate flatness and center-guard gap design and imbalance. A new configuration of center-guard gap with triangular shape cross section has been optimized to obtain the same thermal resistance as a 2 mm wide gap with rectangular shape cross section that has been used in the HTGHPs at NPL and LNE. Recommendations have been made on the selections of heater plate materials, high-temperature high-emissivity coatings and miniature temperature sensors. For the first time, thermal stress analysis method has been applied to the field of HTGHPs, in order to estimate the effect of differential thermal expansion on the flatness of thin rigid specimens during thermal conductivity tests in a GHP.     

Photodecomposition of methylmercury in an Arctic Alaskan lake

Sunlight-induced decomposition of monomethylmercury (MMHg) reduces its availability for accumulation in aquatic food webs. We examined MMHg degradation in epilimnetic waters of Toolik Lake (68 degrees 38' N, 149 degrees 36' W) in arctic Alaska, a region illuminated by sunlight almost continuously during the summer. MMHg decomposition in surface water of Toolik Lake is exclusively abiotic and mediated by sunlight; comparable rates of MMHg decomposition were observed in filter-sterilized and unfiltered surface waters incubated under in situ sunlight and temperature conditions, and no MMHg was degraded in unfiltered aliquots incubated in the dark. Rates of photodecomposition are first order with respect to both MMHg concentration and the intensity of photosynthetically active radiation (PAR), except at the lake surface where rates of photochemical degradation are enhanced relative to PAR intensity and may be attributed to an additional influence of ultraviolet light. The estimated annual loss of MMHg to photodecomposition in Toolik Lake, though limited to a 100-d ice-free season, accounts for about 80% of the MMHg mobilized annually from in situ sedimentary production, the primary source in Toolik Lake. These results suggest that greater light attenuation in lacustrine surface waters, a potential result of increased loadings of dissolved organic matter due to continued warming in the Arctic, may result in less photodecomposition and subsequently greater availability of MMHg for bioaccumulation.     

Effects of inbreeding on production and survival in Holsteins

Responses of registered Holstein cows to various levels of inbreeding were examined with pedigree data supplied by the Holstein Association USA and test-day production data from 1970 through 1998 obtained from the Animal Breeding Center at Cornell University. Rate of increase in level of inbreeding has been accelerating over time, making it more difficult for producers to make matings that avoid the potentially deleterious effects of inbreeding. Milk production losses per lactation caused by inbreeding were generally 35 kg per percentage inbreeding level >0.01 but increased to 55 kg per percentage inbreeding level from 0.07 to 0.10. Somatic cell score was not affected by level of inbreeding. Inbreeding had the greatest effect on production at ages <22 mo and early in lactation. Early onset of the deleterious effects of inbreeding resulted in larger net present value losses than when effects of inbreeding occurred later. Losses were likely enhanced due to the need to freshen animals as early as possible to maximize net present value returns. Survival decreased as level of inbreeding increased and was likely to have a greater negative impact on the financial health of the dairy enterprise than production losses.     

New Transient Hot-Bridge Sensor to Measure Thermal Conductivity, Thermal Diffusivity, and Volumetric Specific Heat

A high sensitivity thermoelectric sensor to measure all relevant thermal transport properties has been developed. This so-called transient hot bridge (THB) decidedly improves the state of the art for transient measurements of the thermal conductivity, thermal diffusivity, and volumetric specific heat. The new sensor is realized as a printed circuit foil of nickel between two polyimide sheets. Its layout consists of four identical strips arranged in parallel and connected for an equal-ratio Wheatstone bridge. At uniform temperature, the bridge is inherently balanced, i.e., no nulling is required prior to a run. An electric current makes the unequally spaced strips establish an inhomogeneous temperature profile that turns the bridge into an unbalanced condition. From then on, the THB produces an offset-free output signal of high sensitivity as a measure of the properties mentioned of the surrounding specimen. The signal is virtually free of thermal emf’s because no external bridge resistors are needed. Each single strip is meander-shaped to give it a higher resistivity and, additionally, segmented into a long and short part to compensate for the end effect. The THB closely meets the specific requirements of industry and research institutes for an easy to handle and accurate low cost sensor. As the key component of an instrument, it allows rapid thermal-conductivity measurements on solid and fluid specimens from 0.02 to 100 W· m⁻¹·K⁻¹ at temperatures up to 250°C. Measurements on some reference materials and thermal insulations are presented. These verify the preliminary estimated uncertainty of 2% in thermal conductivity.     

Geochemical controls on the production and distribution of methylmercury in near-shore marine sediments

We examined temporal differences in sedimentary production of monomethylmercury (MMHg) at three sites in Long Island Sound (LIS). Sediment-phase concentrations of Hg species decreased from west to east in LIS surface sediments, following the trend of organic matter. However, Hg methylation potentials, measured by incubation with an isotopic tracer (200Hg), increased from west to east. 200Hg methylation potentials were enhanced in August relative to March and June, attributable to differences in activity of sulfate-reducing bacteria. Organic matter and acid-volatile sulfide influenced the distribution coefficient (KD) of inorganic Hg (Hg(II) = total Hg - MMHg) and inhibited 200Hg methylation in surface sediments. 200Hg methylation varied inversely with the KD of Hg(II) and positively with the concentration of Hg(II), mostly as HgS0, in LIS pore waters. Accordingly, we posit that a principal control on MMHg production in low-sulfide, coastal marine sediments is partitioning of Hg(II) between particle and dissolved phases, which regulates availability of Hg substrate to methylating bacteria. Most of the partitioning in LIS sediments is due to Hg-organic associations. This suggests that reductions in the organic content of coastal sediment, a potential result of nutrient abatement programs intended to inhibit eutrophication of near-shore waters, could enhance MMHg production by increasing the bioavailability of the large reservoir of "legacy Hg" buried within the sediment.     

Formation of artifact methylmercury during extraction from a sediment reference material.

We examined monomethylmercury (MMHg) in sediment reference material IAEA-405 and sediments from Long Island Sound. MMHg was extracted by both aqueous distillation and leaching with dilute solutions of nitric acid. MMHg was formed from both ambient and added inorganic mercury in extracts of IAEA-405 by either technique. Artifact MMHg in IAEA-405 was related linearly with ambient reactive mercury (HgR) in leachates having > 1.2 M acid, but little or no artifact was measured in less acidic extracts. Addition of potassium chloride enhanced extraction of HgR from LAEA-405 but had no effect on artifact MMHg for each leachate molarity tested. Mercury methylation occurred in solution and was a function of both the availability of HgR and a methylation potential, being limited by HgR in less acidic solutions (0.5-1.2 M) and by the methylation potential in more acidic ones (> 1.2 M). Formation of artifact MMHg in Long Island Sound sediments was inconclusive, but additions of inorganic mercury demonstrated that a potential exists. A potential for abiotic mercury methylation seems always present in sediment, and the availability of HgR appears to control MMHg production under environmental conditions. Abiotic methylation of mercury may occur in environs where the reactivity of inorganic mercury is enhanced, such as river-seawater mixing zones.